Objective:To assess the efficiency of modified enrichment method for cell-free fetal DNA (cffDNA) through purified superparamagnetic beads during non-invasive prenatal testing (NIPT).Methods:A total of 26 252 pregnant women undergoing NIPT at the Maternal and Child Health Care Hospital of Haidian District from December 2017 to September 2022 were recruited and randomly assigned into the conventional group ( n = 10 573) and the modified enrichment group ( n = 15 679), who were then subjected to the screening and enrichment of the cffDNA using a conventional and modified technique, respectively. High-risk pregnant women detected by NIPT were subjected to invasive prenatal diagnosis. All women were followed up for their pregnancy outcomes, and the detection efficacy of the two methods was compared in terms of fragment size, concentration of cffDNA, duplicate detection rate, and indices of clinical laboratory tests. Results:The fragment size of the main peak of the cell-free DNA library of the modified enrichment group was significantly lower than that of the conventional group [267 (264, 269) bp vs. 294 (292, 296) bp, P<0.01], while the concentration of cffDNA was significantly higher [21.86% (17.61%, 26.36%) vs. 9.08% (6.87%, 11.87%), P<0.01]. In addition, the duplicate detection rate (0.740% vs. 2.02%, χ2=83.90, P<0.01) and detection failure rate (0.006% vs. 0.057%, P<0.05) in the modified enrichment group were significantly lower than those of the conventional group. The combined positive predictive value (PPV) in both high-risk (64.3% vs. 76.1%) and low-risk (35.3% vs. 45.5%) pregnant women from the modified enrichment group was slightly lower than those from the conventional group, though no significant difference was detected. There was one false negative case for trisomy 21 among the high-risk pregnant women from the conventional group, and no false negative case was found in the modified enrichment group. Conclusion:The modified technique to screen and enrich the cffDNA has significantly enhanced the relative concentration of cffDNA and reduced the failure and duplication detection rate of NIPT, which has significantly reduced the incidence of false negative cases due to the low concentration of cffDNA, and greatly increased the overall detection efficacy of NIPT.

Objective To explore the trends and patterns of changes in airway inflammation,mitochondrial function,and arginine levels in patients with different degrees of asthma.Methods A total of 122 patients with acute asthma admitted to the Affiliated Traditional Chinese Medicine Hospital of Xinjiang Medical University from November 2016 to May 2017 were selected as the research subjects.According to the severity grading criteria,the patients were divided into the intermittent state group(n=45),mild asthma group(n=23),moderate asthma group(n=30),and severe asthma group(n=24).Another 44 healthy individuals who underwent physical examinations at the Medical Examination Center of the Affiliated Traditional Chinese Medicine Hospital of Xinjiang Medical University from November 2016 to May 2017 were selected as the healthy control group.About 18-20 mL of venous blood was extracted from each group of subjects.The levels of L-arginine(L-Arg)and asymmetric dimethylarginine(ADMA)were detected by high-performance liquid chromatography-tandem mass spectrometry,and the levels of type 1 protein arginine methyltransferase(PRMT1),dimethylarginine dimethylaminohydrolase 1(DDAH1),reactive oxygen species(ROS),thiobarbituric acid reactive substances(TBARS),8-isoprostaglandin(8-iso),lipid peroxide(LPO),and malondialdehyde(MDA)were detected by enzyme-linked immunosorbent assay.Platelets and mitochondria were extracted from each group of subjects.The platelet mitochondrial membrane potential(MMP)was measured using JC-1 flow cytometry,and the mitochondrial adenosine triphosphate(ATP)and cytochrome C oxidase(COX)activity were detected.Results The level of 8-iso in plasma of patients in the intermittent state group,mild asthma group,moderate asthma group,and severe asthma group were significantly higher than that in the healthy control group(P＜0.05);the level of 8-iso in plasma of patients in the severe asthma group was significantly higher than that in the intermittent state group,mild asthma group,and moderate asthma group(P＜0.05);the level of 8-iso in plasma of patients in the intermittent state group,mild asthma group,and moderate asthma group showed an increasing trend,but there was no statistically significant difference(P＞0.05).There was no statistically significant difference in the TBARS level in the plasma of subjects in the five groups(P＞0.05).The levels of ROS and MDA in plasma of patients in the intermittent state group,mild asthma group,moderate asthma group,and severe asthma group were significantly higher than those in the healthy control group(P＜0.05);the levels of ROS and MDA in plasma of patients in the moderate asthma group and severe asthma group were significantly higher than those in the intermittent state group and mild asthma group(P＜0.05);there was no statistically significant difference in plasma ROS and MDA levels between the intermittent state group and the mild asthma group(P＞0.05),and there was no statistically significant difference in plasma ROS and MDA levels between the moderate asthma group and the severe asthma group(P＞0.05).The plasma LPO level of patients in the intermittent state group,mild asthma group,moderate asthma group,and severe asthma group was significantly higher than that in the healthy control group(P＜0.05);the plasma LPO level of patients in the moderate asthma group and severe asthma group was significantly higher than that in the intermittent state group(P＜0.05);there was no statistically significant difference in plasma LPO level of patients in the mild asthma group,moderate asthma group,and severe asthma group(P＞0.05).The mitochondrial MMP,ATP,and COX activity in the intermittent state group,mild asthma group,moderate asthma group,and severe asthma group were significantly lower than those in the healthy control group(P＜0.05);there was no statistically significant difference in mitochondrial MMP,ATP,and COX activity of patients in the intermittent state group,mild asthma group,moderate asthma group,and severe asthma group(P＞0.05).The levels of plasma L-Arg,the ratio of L-Arg/ADMA,and DDAH1 in the intermittent state group,mild asthma group,moderate asthma group,and severe asthma group were significantly lower than those in the healthy control group(P＜0.05);there was no statistically significant difference in plasma L-Arg,the ratio of L-Arg/ADMA,and DDAH1 levels of patients in the intermittent state group,mild asthma group,moderate asthma group,and severe asthma group(P＞0.05).The plasma ADMA and PRMT1 levels showed no statistically significant difference among the five groups of patients(P＞0.05).Conclusion Asthma patients exhibit mitochondrial dysfunction and abnormal arginine metabolism at any stage of the disease.Although there is no statistically significant difference in the change trend at the cellular micro level among patients with different stages of asthma,the trend of changes in oxidative damage in the body is consistent with the aggravation of the disease.

Objective:To evaluate the value of modified magnetic bead screening for enrichment of cell-free fetal DNA (cffDNA) in non-invasive prenatal testing (NIPT).Methods:This study retrospectively recruited 31 cases with low concentration of cffDNA (<6.00%), Z value in the gray zone (3.00-4.00) at the first detection, or false-positive (confirmed by invasive prenatal diagnosis) or false-negative (confirmed by postnatal chromosome test) results among 11 000 pregnant women who underwent routine NIPT in Beijing Haidian District Maternal and Child Health Care Hospital from October 2017 to December 2019. Plasma samples collected for the first-time routine NIPT were used to enrich cffDNA using modified magnetic beads for NIPT (modified NIPT). Wilcoxon rank sum test was used to compare the modified NIPT with the routine NIPT in detecting the cffDNA concentrations of male fetuses.Results:Among the 31 pregnant women, there were 13 cases with low cffDNA concentration in routine NIPT, 11 having false-positive results in the routine NIPT (three for trisomy 13, four for trisomy 18 and four for trisomy 21, all were confirmed by invasive prenatal diagnosis), six with gray-zone Z values in the first-time NIPT (retesting indicating low risk) and one having false negative result for trisomy 21 (confirmed by postnatal chromosome test). Cell-free DNA (cfDNA) fragments less than 150 bp were effectively enriched using the modified magnetic bead screening and the concentration of cffDNA was increased from 4.43% (2.45%-17.61%) in routine NIPT to 13.46% (7.75%-36.64%) in the modified NIPT ( Z=-14.22, P<0.01). Results of the modified NIPT indicated that 13 cases with low cffDNA concentration of routine NIPT were successfully detected as low risk, as well as the risks in the six cases with gray-zone Z value and six of the 11 false-positive cases in the routine NIPT were low, which were consistent with the retest results of the routine NIPT, while high risk was found in one false-negative case. Conclusions:The modified NIPT could reduce the false positive rate by lowering the failure rate caused by low concentration of cffDNA and is able to identify false-negative cases. Compared with the routine NIPT, it shows a higher success rate and a lower false positive rate.

Objective To search for novel potent 3-ester derivatives of arenobufagin and test their antitumor activities in vitro. Methods Target compounds were synthesized by esterification of arenobufagin with acids. CellTiter method was used to assay the in vitro antitumor activities. Results 3-Ester derivatives exhibited excellent antitumor activities against all the cancer cells. Conclusion Among the 3-ester derivatives, compound 2a had the best activities with the IC₅₀ of 4.0−91.7 nmol/L and appeared to be a valuable candidate for further study.

Objective: To evaluate the long-term effect and safety of chrono-chemotherapy combined with intensity modulated radiotherapy (IMRT) in locally advanced nasopharyngeal carcinoma (NPC). Methods: 160 patients with locally advanced NPC were randomly divided into a chrono group and conventional group according to random number table. In the first stage, all patients underwent two cycles of induced chemotherapy, consisting of docetaxel, cisplatin and 5-Fu every 21 days. Notably, patients received chrono-moduated chemotherapy according to circadian rhythm in the chrono group, and conventional chemotherapy in the conventional group. Then, 21 days after the completion of first stage, three cycles of concurrent cisplatin chemotherapy every 21 days were given to all patients during IMRT. The median follow-up after the completion of radiotherapy was 31 months. Long-term side effects and the survival of patients were observed. Results: Patients in the chrono group had significantly lower rates of hearing loss (22.72%), dysphagia (0) and neck fibrosis (4.54%) compared with those in the conventional group (39.13%、8.69%, 15.94%, respectively, all P<0.05). Meanwhile, the 1- year overall survival rates (97.0% vs 92.8%), 3-year overall survival rates (80.3% vs 81.2%), 1-year progression free survival rates (95.5% vs 87.0%), 3-year progression free survival rates (71.2% vs 73.9%), 1-year locoregional relapse-free survival rates (97.0% vs 95.7%), 1-year locoregional relapse-free survival rates (92.4% vs 92.8%), 1-year distant metastasis-free survival rates (97.0% vs 98.6%) and 3-year distant metastasis-free survival rates (90.9% vs 91.3%) between the chrono group and the conventional group were not statistically significant (all P>0.05). Conclusions Compared with conventional chemotherapy, chrono-chemotherapy combined with IMRT didn′t affect long-term survival, but reducing the incidence of adverse events in patients with locally advanced NPC.

Silicon was once considered a biologically inert element, but recent research has shown its value for human health.Soluble silicic acid is the available form of silicon in human body. This paper reviews the absorption, distribution and metabolic characteristics of dietary soluble silicic acid in human body, as well as its relationship with human health.Available data show that it has lots of supporting evidences that dietary soluble silicic acid can prevent osteoporosis, maintain vascular health, improve the symptoms of Alzheimer's disease and multiple sclerosis.Its unique crosslinking ability and antagonism to toxic aluminum play a crucial role. In the early stage of human life, there is a strong demand for silicon, and the level of silicon in the aged is generally reduced, suggesting that pregnant women and old people should pay attention to the intake of soluble silicic acid. It is suggested to strengthen the basic and applied research on dietary soluble silicic acid,and gradually establish the relevant nutrition and hygiene standards.

Silicon was once considered a biologically inert element, but recent research has shown its value for human health.Soluble silicic acid is the available form of silicon in human body. This paper reviews the absorption, distribution and metabolic characteristics of dietary soluble silicic acid in human body, as well as its relationship with human health.Available data show that it has lots of supporting evidences that dietary soluble silicic acid can prevent osteoporosis, maintain vascular health, improve the symptoms of Alzheimer's disease and multiple sclerosis.Its unique crosslinking ability and antagonism to toxic aluminum play a crucial role. In the early stage of human life, there is a strong demand for silicon, and the level of silicon in the aged is generally reduced, suggesting that pregnant women and old people should pay attention to the intake of soluble silicic acid. It is suggested to strengthen the basic and applied research on dietary soluble silicic acid,and gradually establish the relevant nutrition and hygiene standards.

Objective To evaluate the efficacy of 0.3％ sodium hyaluronate ophthalmic solution in mild-to-moderate dry eye patients.Metbods A prospective,multicenter,and self-controlled clinical trial was performed on 200 patients who were diagnosed as mild-to-moderate dry eye from January 2015 to June 2017 in Eye Institute of Xiamen University,Eye & ENT Hospital of Fudan University,Zhongshan Ophthalmic Center,Xiangya Hospital Central South University,Eye Hospital of Wenzhou Medical University,and Beijing Tongren Hospital,Capital Medical University.The study protocol was approved by the Ethics Committees of Eye Institute of Xiamen University,written informed consent was obtained from each patient prior to any medical examination.All patients were treated with 0.3％ sodium hyaluronate ophthalmic solution 6 times per day (one drop each time) for 28 days.Corneal fluorescein sodium staining,tear film break-up time (BUT),Schirmer Ⅰ test (S Ⅰ t),degree of conjunctival hyperemia,eyelid margin,meibomian gland secretion,secretory capacity of meibomian gland and subjective symptoms were assessed at baseline,on the 14th day and 28th day after treatment.Bulbar impression cytology was evaluated at baseline and on the 28th day after treatment.Irritation of 0.3％ sodium hyaluronate ophthalmic solution was estimated on the 14th day and 28th day after treatment.Results The total score of subjective symptoms,BUT,S Ⅰ t,degree of conjunctival hyperemia were significantly different among different treatment time points (F =108.969,27.598,16.838,36.750;all at P＜0.01).Compared with before treatment,the total score of subjective symptoms was significantly decreased,the degree of conjunctival hyperemia and the total corneal fluorescein sodium staining point number were significant decreased on the 14th day and 28th day after treatment.The total score of subjective symptoms,degree of conjunctival hyperemia and total corneal fluorescein sodium staining point number on the 28th day after treatment were significant lower than those on the 14th day after treatment.Compared with before treatment,the BUT was significantly longer and the S Ⅰ t scores were significantly increased on the 14th day and 28th day after treatment.The BUT on the 28th day after treatment was significantly longer than that on the 14th day after treatment;no significant difference in S Ⅰ t was observed between the 28th day and the 14th day after treatment.The scores of palpebral margin change,meibomian gland secretory ability and secretion characteristics were not significantly different among different treatment time points(H=0.255,2.356,0.294;all at P＞0.05).The impression cytology grade on the 28th day after treatment was 1.08±0.74,which was significantly lower than 1.53±0.76 before treatment (t =5.979,P＜0.01).The number of goblet cells on the 28th day after treatment was significantly higher than that before treatment (U =1 806.500,P＜ 0.01).On the 14th day after treatment,70％ of the patients indicated that the drug was non-irritating,and no patient had intolerable irritation affecting daily lives.All patients had good tolerance to this drug.Conclusions The use of 0.3％ sodium hyaluronate eye drops can improve the symptoms and signs of mild to moderate dry eye,which can be widely used for mild-to-moderate dry eye patients in clinic.

Objective： ：To study the effect of silencing DKK1 (Dickkopf1) gene on the proliferation, cell cycle and apoptosis of gastric cancer AGS cells and the action mechanism. Methods： ：The DKK1-shRNA vector was constructed and transfected into AGS cells. The stably transfected cell lines were screened. The total protein and RNAof the transfected cells were extracted and the mRNAand protein expressions of DKK1 were detected by qPCR and WB, respectively. The experiment was divided into blank control group (Control), negative control group (shNC) and DKK1 silence group (DKK1-shRNA). CCK8 assay was used to detect the proliferation ofAGS cells of each group cultured for 0, 24, 48, 72, 96, 120 and 144 h, and flow cytometry was used to analyze the cell cycle and apoptosis in each group. The relationship between DKK1 and clinicopathological features of gastric cancer was analyzed after searching HPA database. Results：The gastric cancer AGS cells with stable DKK1 gene knockdown was successfully established, and it was confirmed that the mRNA and proteinexpressions of DKK1 in DKK1-shRNA group decreased by 72% and 47%, respectively, compared to shNC group (all P<0.05). The cell proliferation curve showed that, the cell proliferation in DKKl-shRNAgroup significantly decreased after 72 hour of culture compared with that in control and shNC groups (P<0.05). The cell number of S phase decreased from 32.06% to 25.87%, while the number of G2/M phase increased from 8.49% to 21.26% compared with shNC group (all P<0.05). The number of apoptotic cells also statistically increased from 10.34% to 20.65% (all P<0.05). The data of HPAdatabase showed that DKK1 mRNAlevel in gastric cancer tissues was significantly higher than that in normal tissues, and the high expression of DKK1 mRNAwas negatively correlat ed with the survival rate of gastric cancer patients. Conclusion: : Silencing DKK1 gene can inhibit the proliferation of gastric cancer cells, arrest cells in G2/M phase and promote cell apoptosis. DKK1 plays a pro-carcinogenic effect in gastric cancer.

Objective To investigate the efficacy,adverse reactions and immune function of time-adjusted chemotherapy combined with intensity-modulated radiation therapy (IMRT) and conventional chemotherapy combined with IMRT for locally advanced nasopharyngeal carcinoma.Methods Random number grouping method was used to divide 66 cases of locally advanced nasopharyngeal carcinoma into 2 groups,of which 36 cases in the time-adjusted chemotherapy group and 30 cases in the conventional group.Both of them received docetaxel + cisplatin + fluorouracil regimen to induce chemotherapy for 2 cycles.The time-adjusted chemotherapy group was treated with intravenous injection of electronic automatic injection pump,the conventional group was treated with conventional intravenous infusion,and both groups were treated with synchronous cisplatin combined with IMRT.Calculated survival rate was generated by Kaplan-Meier method and long-term adverse reactions was evaluated according to CTC 3.0 criteria.Results The 3-year overall survival (OS) rate was 86.1％ and 93.3％ in the time-adjusted chemotherapy group and the regular group,the 3-year progress-free survival (PFS) was 83.3％ and 93.3％,the 3-year RFS was 88.5％ and 93.3％,and the 3-year recurrence-free survival was 94.1％ and 100％ respectively with no statistically significant difference (P ＞ 0.05).The dryness and hearing loss of the time-adjusted chemotherapy group had a decreasing trend compared with the conventional group.However,CD3 +,CD3 + CD4 +,CD3 + CD4 + CD8 +,and CD4 +/CD8 + of the time-adjusted chemotherapy group had an increasing trend compared with the conventional group.Conclusions Both time-adjusted chemotherapy and conventional chemotherapy combined with IMRT had comparable mid-term efficacy,but the former had lower adverse reactions,improved quality of life and immune function.Trial registration Chinese clinical trial registry,ChiCTR1800016809